A comprehensive study on the contents and leaching of trace elements from fly-ash originating from polish hard coal by NAA and AAS methods

In order to assess the environmental risks associated with the emission of fly-ash into the atmosphere and its storage on waste heaps, the trace element contents of fly-ashes from burning Polish hard coal were determined by a newly developed INAA method. Leaching of trace elements from the fly-ash by water and H₂SO₄ solution (pH≈2.5) simulating acid rain, respectively, was studied using AAS and spectrophotometric methods. Analogous experiments were done with neutron-irradiated fly-ash, following the composition of the eluate gamma-spectrometrically. The new fine fly-ash (CTA-FFA-1) candidate reference material was prepared, and the certification was undertaken on the basis of an international intercomparison run. Preliminary evaluation of results shows that at least 38 elements will be certified and, in addition, the “information values” for at least 12 elements will be given.     

Trace elemental content of biological materials

The advantages and disadvantages of neutron activation analysis (NAA) and inductively coupled plasma-source mass spectrometry (ICP-MS) for the analysis of biological materials is reviewed. Comparison is made between NAA (instrumental) and ICP-MS (conventional pneumatic solution nebulization and laser ablation) analysis of the biological reference material National Bureau of Standards (NBS) SRM 1577 Bovine Liver. Relatively good agreement is achieved between the results for the 18 elements analyzed by both techniques and those either certified or reported in the literature. Elemental concentrations for Li, Mg, Al, Ca, Cr, Mn, Fe, Cu, Zn, Br, Rb, and Cs are also reported for IAEA Mixed Human Diet (H9), NBS SRM 909 Human Serum, and NBS SRM 1577a Bovine Liver, analyzed by solution nebulization ICP-MS.     

Determination of mercury in human serum and packed blood cells by neutron activation analysis

A method is described for the determination of mercury in human blood serum and packed blood cells employing neutron activation analysis. Great attention was devoted to the collection and manipulation of the samples. The accuracy and precision of the method were tested by analyzing biological reference materials and by comparing the concentrations measured in a number of serum samples to those obtained by another, independent technique (cold vapor atomic absorption spectrometry) in the same samples. The article reports the levels measured in blood serum and packed blood cells samples from 15 adult volunteers, as well as the figures determined in a “second-generation” biological reference material (freeze-dried human serum), prepared and conditioned at the University of Ghent.     

An innovative LC/MS approach applied to the determination of zeralenone in maize: Alternate Isotope-coded Derivatization Assay (AIDA)

Zearalenone is a mycotoxin mainly produced by severalFusarium species, which are known to colonize grains in temperate climates. The purpose of the study is to provide a reliable isotope dilution method for the quantification of this mycotoxin. A derivative of the analyte to be used as standard is obtained by reaction with acetic anhydride, which is available in two pure isotopic forms, a protonated (“light”) and a hexadeuterated (“heavy”). The derivatized standards are added to the matrix split intwo parts. Then, the derivatization procedure is repeated on both matrices derivatizing the part containing the “heavy” labelled standard with the “light” acetic anhydride and the part containing the “light” labelled standard with the “heavy” acetic anhydride. Both extracted mixtures are analyzed by LC/MS, monitoring the “light” and the “heavy” labelled analytes and using the former as standard for the latter in one case and viceversa in the other case. The method allowed to obtain very good results, without the need of IAC purification. Presented at the 27^th Mykotoxin-Workshop, Dortmund, Germany, June 13–15, 2005. Financial support: The Italian Ministry of Health     

Certified reference materials (GBW09170 and 09171) of creatinine in human serum

Creatinine is the most widely used clinical marker for assessing renal function. Concentrations of creatinine in human serum need to be carefully checked in order to ensure accurate diagnosis of renal function. Therefore, development of certified reference materials (CRMs) of creatinine in serum is of increasing importance. In this study, two new CRMs (Nos. GBW09170 and 09171) for creatinine in human serum have been developed. They were prepared with mixtures of several dozens of healthy people's and kidney disease patient's serum, respectively. The certified values of 8.10, 34.1 mg/kg for these two CRMs have been assigned by liquid chromatography-isotope dilution mass spectrometry (LC-IDMS) method which was validated by using standard reference material (SRM) of SRM909b (a reference material obtained from National Institute of Standards and Technology, NIST). The expanded uncertainties of certified values for low and high concentrations were estimated to be 1.2 and 1.1%, respectively. The certified values were further confirmed by an international intercomparison for the determination of creatinine in human serum (Consultative Committee for Amount of Substance, CCQM) of K80 (CCQM-K80). These new CRMs of creatinine in human serum pool are totally native without additional creatinine spiked for enrichment. These new CRMs are capable of validating routine clinical methods for ensuring accuracy, reliability and comparability of analytical results from different clinical laboratories. They can also be used for instrument validation, development of secondary reference materials, and evaluating the accuracy of high order clinical methods for the determination of creatinine in human serum.     

A “Nonsterile” Method for Selecting and Growing <Emphasis Type="Italic">Arabidopsis thaliana</Emphasis> Transformants (T<Subscript>2</Subscript> Transgenic Lines) Resistant to Kanamycin

A breakthrough in transgenic Arabidopsis thaliana research was the development of the floral dip transformation protocol, a simple and reliable method of obtaining transformants, T₁ transgenic lines, at high efficiency while avoiding the use of tissue culture. However, the traditional protocol (a “sterile” method) of obtaining T₂ transgenic lines has not evolved along with improvements in transformation technology as it continues to be laborious and time-consuming. In this study, we report on the development of an improved protocol (a “nonsterile” method) for selecting and growing A. thaliana transformants (T₂ transgenic lines) resistant to kanamycin under nonsterile conditions. This protocol involves the use of a simple yet specialized device that will aid in solium selection of T₂ transgenic lines that can be rapidly grown in a hydroponic system. The “nonsterile” method reduces labor and time involved as compared to the “sterile” method; moreover, it is easy to set up and maintain. This method may also be applicable to other selecting agents, and perhaps to other plants.     

The effects of external potassium and magnesium concentrations on the magnesium and potassium inflow rates and growth of micropropagated cherry rootstocks, “F.12/1” (Prunus avium L.) and “Colt” (Prunus avium L.) × Prunus pseudocerasus L.)

The effects (and interaction) of two solution concentrations of Mg (50, 500, μM) and two of K (250, 4250 μM) on the growth of micropropagated plants of “F. 12/1” and “Colt” were investigated using a flowing solution culture system. Magnesium inflow and growth of “Colt” and “F. 12/1” were inhibited to a similar extent by an increased concentration of K in the nutrient solution. However, the consequences of this inhibition were different. Reduced inflow of Mg in “F. 12/1” caused Mg deficiency symptoms at high and low concentrations of K, whereas this only occurred with a combination of high K concentration and low Mg concentration in “Colt”. The distribution of dry matter within the plant was significant in determining susceptibility to Mg deficiency. Since “F. 12/1” has a smaller root:shoot ratio than Colt it is unable to sustain the same concentration of Mg in leaves as “Colt” irrespective of external K concentration. The molar ratio of K:Mg in soil solutions should remain <8.5:1 in order to ensure maximum growth of “F. 12/1” and “Colt”. This revised version was published online in June 2006 with corrections to the Cover Date.     

Modeling of the actomyosin ATPase activity

In the rapid “quench” kientics of myosin, the “initial phosphate burst” is the excess inorganic phosphate that is produced during the early time-course of ATP hydrolysis by myosin subfragment-1 (S-1) or HMM. In general, the existence of a Pi burst implies a rapid (i.e., generally an order of magnitude faster than the steady-state hydrolysis rate) lysis of the phospho-anhydride bond within the ATP molecule, followed by one or more slower steps that are rate limiting for the process. Thus, the presence of a Pi burst can provide an important clue to the mechanism of the reaction. However, in the case of actomyosin, this clue as long been the subject of controversy and misunderstanding. To measure the (initial) Pi burst, myosin S-1 (or HMM) is rapidly mixed with ATP and then the mixture is acid quenched after a specific time period. The medium produced contains free Pi generated from hydrolysis of the ATP. The quantitative measure of the phosphate generated in this way has always been significantly greater than that expected by steady-state “release” of Pi alone, and it is that very difference between this measured Pi after the quench and that amount of Pi expected to be released by steady-state considerations in that same time period that has been referred to as the “initial Pi burst”. Recent investigations of the kinetics of Pi release have used an entirely new method that directly measures the release of Pi from the enzyme-product complex. These studies have made reference to the properties of the “initial Pi burst” in the presence of actin, as well as to a new kinetic entity: the “burst of Pi release”, and have been often vague concerning the true nature of the initial Pi burst, as well as the properties of Pi release as predicted by the current models of the actin activation of the myosin ATPase activity. The purpose of the current article is to correct this oversight, to discuss the “burst” in some detail, and to display the kinetics predicted by the current models for the actin activation of myosin. Furthermore, predictions for the kinetics of the new “burst of Pi release” are discussed in terms of its ability to discriminate between the two current competing models for actin activation of the myosin ATPase activity.     

Md-ACS1 and Md-ACO1 genotyping of apple (<Emphasis Type="Italic">Malus</Emphasis> x <Emphasis Type="Italic">domestica</Emphasis> Borkh.) breeding parents and suitability for marker-assisted selection

Fruit ethylene production genotypes for Md-ACS1 and Md-ACO1 were determined for 60 apple cultivars and 35 advanced breeding selections. Two alleles for each gene are commonly found in cultivated apple. Earlier studies showed that genotypes homozygous for the ACS1-2 allele produce less ethylene and have firmer fruit than ACS1-1/2 and ACS1-1/1 genotypes. ACO1 plays a minor role compared to ACS1, with homozygous ACO1-1 having lower ethylene production. In this study, ACS1-2 and ACO1-1 homozygotes had firmer fruit at harvest and after 60 days of 0–1°C cold storage compared to other genotypes. These genotypes, ACS1-2/2 and ACO1-1/1, were observed for the following 8 of 95 cultivars/selections: “Delblush”, “Fuji”, “Pacific Beauty”, “Sabina” and four breeding selections. Cultivars/selections that were homozygous ACS1-2 but not ACO1-1 were: “Ambrosia”, “Aurora Golden Gala”, “CrimsonCrisp”, “Gala”, “GoldRush”, “Huaguan”, “Pacific Rose, “Pacific Queen”, “Pinova”, “Sansa”, “Sonja”, “Sundance”, “Zestar”, and 17 breeding selections. Cultivars with the heterozygous ACS1-1/2 genotype were “Arlet”, “Braeburn”, “Cameo”, “Delicious”, “Delorgue”, “Empire”, “Enterprise”, “Ginger Gold”, “Golden Delicious”, “Granny Smith”, “Honeycrisp”, “Orin”, “Pink Lady”, “Silken”, “Suncrisp”, “Sundowner”, “Sunrise” and 11 breeding selections. No cultivars were detected homozygous for both ACS1-1 and ACO1-1, or for both ACS1-2 and ACO1-2. This study is the first large-scale allelic genotyping of both ethylene synthesis genes for a comprehensive set of apple breeding parents used in an ongoing breeding project. The data reported here are important for informative selection of parent combinations and marker-assisted selection of progeny for breeding low ethylene-producing apple cultivars for better storability and improved consumer acceptance.     

IAEA internet database of natural matrix reference materials

The International Atomic Energy Agency maintains a database on internationally available certified reference materials of natural origin. The database was updated in 1998 and prepared for an Internet implementation. A user-friendly structure was created, providing two main pathways for browsing, either according to the matrix classification or the producer’s name. The database presently contains over 20,000 values for 480 measurands and 1085 reference materials from 43 different producers. Most of the materials entered contain values for trace and minor elements (66%).     

On the stochastic interpretation of a pathological cell renewal system,I: Theoretical basis

The population constituted by parenchymal liver cells is normally a static or slowly expanding one. This is in contrast with “renewal systems,” or cell populations which turn over more or less rapidly. In experimental situations tending to produce cirrhosis of the liver the rate of cell renewal is appreciable. The situation then corresponds to the definition of a “renewal system.” A repeating unit of the liver lattice structure is proposed in which liver cells are assumed to be sequestrated by anatomical constraints on cell migration, and the order of magnitude of its population determined. The characteristics of the simple birth-and-death process are reviewed to show how it leads to greater fluctuation than does a pure birth process with equivalent expected net growth. A hypothetical birth-and-death process is proposed for the renewal of the cell population of the liver unit. The rate of cell proliferation is placed at a level comparable to that found in reports of experimental cirrhosis taken from the literature. This example of a birth-and-death process leads to the prediction of appreciable fluctuation in the population of the liver unit. It is suggested that fluctuations of this kind may account for some of the morphological features of cirrhosis and lead to a new definition of “nodular regeneration.”     

Apomorphy-based definition also pinpoints a node, and PhyloCode names prevent effective communication

Acceptable methods of defining taxon (or clade) names in the draft PhyloCode, or so-called phylogenetic nomenclature, are “node based,” “stem based,” and “apomorphy based.” All of them define a clade name by pinpointing a node; whereas node-based and stem-based definitions require two or more taxon “specifiers” to define names, an apomorphy-based definition requires two specifiers of different types; namely, a single-taxon specifier and a character specifier. The taxon specifier in an apomorphy-based definition is completely different from the “type” in the Linnaean system. Taxon (or clade) names in the PhyloCode are characterized in two entirely different manners: One is a name that does not change, either in its orthography or in the contents of the taxon referred to by it (or its meaning) over time; the other is a name that is just like a pure mark and thus has no meaning. Communication through such PhyloCode names is very ineffective or impossible.     

Detection of gene flow from GM to non-GM watermelon in a field trial

Gene flow from genetically modified (GM) crops to conventional non-GM crops is a serious concern for protection of conventional and organic farming. Gene flow from GM watermelon developed for rootstock use, containing cucumber green mottle mosaic virus (CGMMV)-coat protein (CP) gene, to a non-GM isogenic control variety “Clhalteok” and grafted watermelon “Keumcheon” was investigated in a small scale field trial as a pilot study. Hybrids between GM and non-GM watermelons were screened from 1304 “Chalteok” seeds and 856 “Keumcheon” seeds using the duplex PCR method targeting theCGMMV- CP gene as a marker. Hybrids were found in all pollen recipient plots. The gene flow frequencies were greater for “Chaiteok” than for “KeumcheonD; with 75% outcrossing in the “Chaiteok” plot at the closest distance (0.8 m) to the GM plot. A much larger scale field trial is necessary to identify the isolation distance between GM and non-GM watermelon, as the behaviors of insect pollinators needs to be clarified in Korea.     

Determination of trace elements in human serum by inductively coupled plasma-mass spectrometry

The determination of trace and ultratrace elements in human serum by ICP-MS is described. The accuracy of the method is tested using a “second generation” human serum reference material. Elements determined include Fe, Co, Cu, Zn, Br, Rb, Sr, Mo, and Cs. The method is compared to nuclear analytical methods (NAA, PIXE). Perspectives for the future are also discussed.     

Two types of using of space in the resident common shrews <Emphasis Type="Italic">Sorex araneus</Emphasis> L.

The home range of resident animals is considered as “familiar area” including a “foraging area.” It has been revealed that the activity of an average animal unit in the “foraging area” could be approximated by normal distribution. Estimation of activity distribution in the “familiar area” (beyond the “foraging area”) was impeded by means of marking since it might be difficult to record distant movements, and the method does not provide an essential body of data. In the case of the common shrew Sorex araneus, the “familiar area” was estimated using pitfall as animals evade them in the known areal. The “foraging area” radius of the average shrew was taken to be 30 m (95% of the animal unit activity), the radius of “familiar area” was within the range of 180–240 m. The “foraging area” was expected to provide the animal with vital resources, and the “familiar area” reflects its need for exploratory activity.     

Iota-carrageenan from a newly farmed, rare variety of eucheumoid seaweed—“endong”

A seaweed with the local name “endong” is a rare eucheumoid variety that has recently been farmed in Tawi-Tawi, southern Philippines. Local farmers describe it as “eel-like” because of its long, slender and smooth thalli. Being somewhat similar in appearance to the dominantly farmed seaweed “tambalang” (Kappaphycus alvarezii var. tambalangii), farmers mix their harvests of “endong” with those of “tambalang”. We analyzed the hydrocolloid extract from “endong” using spectroscopic and chemical techniques. The extract’s infrared and nuclear magnetic resonance (¹H and ¹³C) spectra, and sulfate and 3,6-anhydrogalactose contents revealed carrageenan of the iota-type, similar to extracts from another farmed species, Eucheuma denticulatum. This result implies that “endong” carrageenan exhibits physico-chemical properties different from those of kappa-carrageenan of “tambalang”. The findings of this study are of significance to the seaweed industry, as carrageenan quality problems could potentially arise from a mixed harvest. Seaweed farmers are advised to separate their harvests of “endong” and “tambalang”. Presented at the 19th International Seaweed Symposium, Kobe, Japan     

Cascade Impactor (CI) Mensuration—An Assessment of the Accuracy and Precision of Commercially Available Optical Measurement Systems

Multi-stage cascade impactors (CIs) are the preferred measurement technique for characterizing the aerodynamic particle size distribution of an inhalable aerosol. Stage mensuration is the recommended pharmacopeial method for monitoring CI “fitness for purpose” within a GxP environment. The Impactor Sub-Team of the European Pharmaceutical Aerosol Group has undertaken an inter-laboratory study to assess both the precision and accuracy of a range of makes and models of instruments currently used for optical inspection of impactor stages. Measurement of two Andersen 8-stage ‘non-viable’ cascade impactor “reference” stages that were representative of jet sizes for this instrument type (stages 2 and 7) confirmed that all instruments evaluated were capable of reproducible jet measurement, with the overall capability being within the current pharmacopeial stage specifications for both stages. In the assessment of absolute accuracy, small, but consistent differences (ca. 0.6% of the certified value) observed between ‘dots’ and ‘spots’ of a calibrated chromium-plated reticule were observed, most likely the result of treatment of partially lit pixels along the circumference of this calibration standard. Measurements of three certified ring gauges, the smallest having a nominal diameter of 1.0 mm, were consistent with the observation where treatment of partially illuminated pixels at the periphery of the projected image can result in undersizing. However, the bias was less than 1% of the certified diameter. The optical inspection instruments evaluated are fully capable of confirming cascade impactor suitability in accordance with pharmacopeial practice.     

Decline in energy storage in the Antarctic minke whale (<Emphasis Type="Italic">Balaenoptera bonaerensis</Emphasis>) in the Southern Ocean

The annual trend in energy storage in the Antarctic minke whale was examined using catch data from all 18 survey years in the Japanese Whale Research Program (JARPA). Regression analyses clearly showed that blubber thickness, girth and fat weight have been decreasing for nearly 2 decades. The decrease per year is estimated at approximately 0.02 cm for mid-lateral blubber thickness and 17 kg for fat weight, corresponding to 9% for both measurements over the 18-year period. Furthermore, “date”, “extent of diatom adhesion”, “sex”, “body length”, “fetus length”, “latitude”, “age” and “longitude” were all identified as partially independent predictors of blubber thickness. The direct interpretation of this substantial decline in energy storage in terms of food availability is difficult, since no long-term krill abundance series is available. However, an increase in the abundance of krill feeders other than minke whales and a resulting decrease in the krill population must be considered as a likely explanation.     

Ribulose-1,5-bisphosphate carboxylase/oxygenase from thermophilic cyanobacterium Thermosynechococcus elongatus

Ribulose-1,5-bisphosphate carboxylase/oxygenase (rubisco) can be divided into two branches: the “red-like type” of marine algae and the “green-like type” of cyanobacteria, green algae, and higher plants. We found that the “green-like type” rubisco from the thermophilic cyanobacterium Thermosynechococcus elongatus has an almost 2-fold higher specificity factor compared with rubiscos of mesophilic cyanobacteria, reaching the values of higher plants, and simultaneously revealing an improvement in enzyme thermostability. The difference in the activation energies at the transition stages between the oxygenase and carboxylase reactions for Thermosynechococcus elongatus rubisco is very close to that of Galdieria partita and significantly higher than that of spinach. This is the first characterization of a “green-like type” rubisco from thermophilic organism.